There was a need to suppress three genes (tKD) at once in tumor cells. According to the literature, there are no problems with triple (tKO) or even quadruple (qKO) gene knockouts. However, we do not have the opportunity to carry out a gene KO, and there is no information on knockdown using more than two siRNA. Could you tell me, please, whether the knockout of three genes will work at the same time and what problems can there be?
Thank you!
My only experience in triple KD is using a particular brand of siRNAs: siPOOLs (https://www.sitoolsbiotech.com/sipools.php) which according to the manufacturer are specifically designed to lower the amount of off target effects. Using them, I managed to get triple knockdowns with no visible side effects on the cells except maybe an increased cell mortality, which is anyway expected I guess when you are altering the expression of 3 genes. You would need to adjust the protocols to ensure you have enough lipofectamine or whatever carrier you use for your siRNAs, and of course increase the concentration of your negative control siRNAs to match the concentration of the combined target siRNAs, but other than that it should be pretty straightforward.
I must say I never did an RNAseq of this triple KD so I don't know if there are increased off target effects, but I would suspect there may be some.
- Badges
- Science topic
- Similar topics
- Biological Science
- Microbiology
- Virology
- Virus